An SSR reaction system for <em>Abies beshanzuensis</em> progeny with artificial pollination and progeny identification

LIN Er-pei; MA Hai-quan; FAN Min-liang; LUO Wen-jian; HUANG Hua-hong; TONG Zai-kang

doi:10.11833/j.issn.2095-0756.2011.02.010

Volume 28 Issue 2

Apr. 2011

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Article Navigation > Journal of Zhejiang A&F University > 2011 > 28(2): 234-240

LIN Er-pei, MA Hai-quan, FAN Min-liang, LUO Wen-jian, HUANG Hua-hong, TONG Zai-kang. An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification[J]. Journal of Zhejiang A&F University, 2011, 28(2): 234-240. doi: 10.11833/j.issn.2095-0756.2011.02.010

Citation:

LIN Er-pei, MA Hai-quan, FAN Min-liang, LUO Wen-jian, HUANG Hua-hong, TONG Zai-kang. An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification[J]. Journal of Zhejiang A&F University, 2011, 28(2): 234-240. doi: 10.11833/j.issn.2095-0756.2011.02.010

An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

doi: 10.11833/j.issn.2095-0756.2011.02.010

1.
The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University，Lin’an 311300，Zhejiang，China;
2.
Zhejiang Qingyuan Forestry Enterprise，Qingyuan 323800，Zhejiang，China;
3.
Forest Tree Seed and Stock Master Station of Zhejiang Province，Hangzhou 310020，Zhejiang，China

More Information

Corresponding author: HUANG Hua-hong
Publish Date: 2011-04-20

Abstract

To establish a Simple Sequence Repeat with Polymerase Chain Reaction （SSR-PCR） system for Abies beshanzuensis and to optimize the factors which affect the PCR reaction，the cetyl-trimethyl-ammonium bromide （CTAB） method was used to extract DNA from its leaves. Then，to evaluate the universality of the primers，the amplified product of one pair of primers were cloned and sequenced. Finally，primer pairs with good amplification and diversity were selected. Results showed that the optimal PCR reaction system was 60 ng template DNA，3.75 mmolL－1 Mg2+，0.15 mmolL－1 dNTPs，0.4 molL－1 primers，and 16.67 nkat （1.0 U） Taq polymerase in a volume of 20 L. Blast results also showed that the identity value reached 94%. Then，15 pairs of primers with good amplification and diversity were obtained with four that could produce differing DNA fragments between Abies beshanzuensis and Abies firma. Identity value results indicated that SSR primers from other Taxodiaceae species could be used with Abies beshanzuensis；additionally the SSR-PCR technique was applicable to preliminary identification of progeny from artificial pollination.［Ch，7 fig. 2 tab. 16 ref.］
- forest tree breeding,
- SSR marker,
- reaction system,
- Abies beshanzuensis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University，Lin’an 311300，Zhejiang，China;

2. Zhejiang Qingyuan Forestry Enterprise，Qingyuan 323800，Zhejiang，China;

3. Forest Tree Seed and Stock Master Station of Zhejiang Province，Hangzhou 310020，Zhejiang，China

Corresponding author: HUANG Hua-hong

Publish Date: 2011-04-20

Key words:

Abstract: To establish a Simple Sequence Repeat with Polymerase Chain Reaction （SSR-PCR） system for Abies beshanzuensis and to optimize the factors which affect the PCR reaction，the cetyl-trimethyl-ammonium bromide （CTAB） method was used to extract DNA from its leaves. Then，to evaluate the universality of the primers，the amplified product of one pair of primers were cloned and sequenced. Finally，primer pairs with good amplification and diversity were selected. Results showed that the optimal PCR reaction system was 60 ng template DNA，3.75 mmolL－1 Mg2+，0.15 mmolL－1 dNTPs，0.4 molL－1 primers，and 16.67 nkat （1.0 U） Taq polymerase in a volume of 20 L. Blast results also showed that the identity value reached 94%. Then，15 pairs of primers with good amplification and diversity were obtained with four that could produce differing DNA fragments between Abies beshanzuensis and Abies firma. Identity value results indicated that SSR primers from other Taxodiaceae species could be used with Abies beshanzuensis；additionally the SSR-PCR technique was applicable to preliminary identification of progeny from artificial pollination.［Ch，7 fig. 2 tab. 16 ref.］

An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

doi: 10.11833/j.issn.2095-0756.2011.02.010

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An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

doi: 10.11833/j.issn.2095-0756.2011.02.010

Corresponding author: HUANG Hua-hong

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An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

doi: 10.11833/j.issn.2095-0756.2011.02.010

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An SSR reaction system for Abies beshanzuensis progeny with artificial pollination and progeny identification

doi: 10.11833/j.issn.2095-0756.2011.02.010

Corresponding author: HUANG Hua-hong

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